This project focused on developing the techniques for the single-cell secretion dynamics measurement, which will contribute to understanding of the detailed mechanism of Immune systems reguration at single-cell resolution. During this project, I succeeded in developing a novel micro fabricated waveguide chip for wide field total internal reflection fluorescence microscopy (TIRFM).This chip has the optical refraction index matching layer on both side of quartz light wave guide slide, and nano-litter well array made by PDMS on top side.Using this chip with the wave-guide illuminator, we measured single-cell cytokine secretion dynamics from hundreds of single-cells every minute for over 2hours.This optical wave-guide based TIRFM system has great potential for revealing single-cell secretion dynamics and will contribute to develop the measurement technique for analyzing cell-cell interaction via humoral factors.